Figure 1. Turtle ready for dissection. Dotted lines indicate incision
sites for removal of plastron and brain exposure.

16



An example is the diagnosis of drowning. Certain classical signs of
drowning are present in warm blooded animals that have also been observed
in sea turtles. However, there is no certainty with present knowledge that
they are diagnostic of drowning in sea turtles.

ACKNOWLEDGMENTS

We thank R. Shoop, R. Prescott and L. Ehrhart for supplying specimens
and T. Murphy, C. Ruckdeschel and M. Lutcavage for their advice and B.
Watkins for the illustrations. A. Carr, D. Odell, M. McGehee, J. Leong,
T. Murphy, G. Zug, A. Rhodin, H. Kumpf, W. Pringle, F. Berry, and S.
Lincoln reviewed and made excellent suggestions on the draft manuscript.

REFERENCES

Ashley, L.M. 1955. Laboratory anatomy of the turtle. Wm. C. Brown Co.,
Dubuque, lowa. 48 p.

Jacobson, E.R. 1978. Reptile necropsy protocol. Journal of Zoo Animal
Medicine. 9:7-13.

‘Van Kruningen, H.J. 1971. Veterinary autopsy procedure. Veterinary
Clinics of North America. 1(1):163-189.


ystem removed revealing

and genitourinary system (16-19).

Liver, head, and gastrointestinal s
lungs (15)

Figure 5.

20



INTRODUCTION

This manual is a guide for necropsies conducted on sea turtles under
ideal laboratory conditions. Since most sea turtle necropsies will be
conducted in the field, the manual should serve primarily as a guide.
However, the manual contains information to conduct a careful and exacting
necropsy, when it is possible to do so.

The value of a necropsy is dependent on the freshness of. the cadaver
and the number of observations. Field biologists are often pressed for
time but wish as much information as possible. Unfortunately, there are
no short cuts. Under these circumstances it would be best to remove the
plastron and take samples for fixation from as many organs as possible.

Necropsies should always be conducted so that a minimum of logistical
problems will occur. The prosector (person conducting necropsy) should
dress in light, easily washable clothes and wear gloves. If possible,

a shaded, well ventilated, and easily cleanable site should be used.
The site should also be chosen in relation to carcass disposal. Large
animals are very difficult to incinerate and must be buried. Disposal
should be in compliance with authorization from proper law enforcement
agents or with a valid state or federal sea turtle permit.

Descriptions are based on the anatomy of the loggerhead sea turtle,
Caretta caretta. Left and right as used in this manual refer to the
turtle's left and right. All measurements should be made in metric units.

EXTERNAL EXAMINATION

The sea turtle's weight, carapace length, carapace width and plastron
length are recorded; note whether straight line or curved measurements
are taken; straight line measurements are preferred. The general condition
of the body is noted, because it reflects the state of decomposition and
nutrition. Decomposition or postmortem autolysis is recognized by color
changes of the skin, collection of blood in dependent areas (rigor mortis;
Stage 1), and by the production of gas a foul odor (putrefaction;
Stage 2). Within a short time after death, the ventral skin becomes
pink to purple and as postmortem changes progress, frank lines of
purple-green will be seen encircling the neck and limbs.

If the carcass is bloated so that gas escapes when a knife is inserted,
advanced decomposition has occurred. At this point, the turtle has a foul
odor. In such cases and in those instances in which the flesh is falling
from the skeletal system, necropsy methods are of little value, because
postmortem changes will mask antemortem events. The determination of
the degree of postmortom decomposition, and therefore the value of necropsy,
is subjective and learned by experience.

Nutritional state can be determined by both external and internal
examination. A starving or chronically i1] turtle has markedly sunken
eyes and plastron. Its skin, carapace and plastron may be covered by an


Figure 2. Muscle masses present after plastron removal. Dotted lines
indicate incision sites to expose viscera.

17



Figure 4, Viscera reflected to show location of spleen (13) and gall
bladder (14), and points of gastrointestinal ties to prevent
loss of stomach contents (N,0,P).

1g



Date Submitted PATHOLOGY RECORD Accession No.
MARINE PATHOLOGY LABORATORY
COLLEGE OF RESOURCE DEVELOPMENT
UNIVERSITY OF RHODE ISLAND
KINGSTON, R. 1. 02881

Veterinarian Address Phone

Owner Address

Ident. Genus Species Length Age Sex Weight Prev. Acces.

Clinical Diagnosis
History and Clinical Summary:
Specimen Submitted Preservation Condition of Specimen
Live Animal] Dead Animal C] | Fresh C] — Frozen] ~— Fixed [1]_| When Received at Lab:
Tissues C]

Autopsy Data Mode of Euthanasia Time and Date of Death Time and Date

Natural Death ~=(] of Autopsy

Tissues Submitted:

Findings:

Cause of Death:

Diagnoses Classification Tissues saved =]
Photographs (_]
Radiographs [7]

Pathologist Date Prepared Fee

White — ORIGINATOR

Yellow — VETERINARIAN

Pink — PATHOLOGIST



of the bladder and cloaca anteriorly. The testes and epididymides occupy
the same area as the ovaries and their ducts.

The interrenal and chromaffin tissues (adrenal cortex and medulla)
are not discrete and are on the ventral surface of the kidney. It is wise
therefore to use wide excision when removing the kidneys and to save renal
peritoneum for location of these structures.

Vv. Connective Tissue, Skeletal Muscle, and Osseous System

The major pectoral and pelvic muscle masses should be carefully
palpated for evidence of mass lesions, and if necessary, incised and
examined. The shoulder and hip joint should be opened and articular
cartilaginous surfaces and synovial fluid examined, noting the color and
consistency of both cartilage and fluid. At least one front flipper and
one rear flipper should be disarticulated at the shoulder and hip joint
and removed in toto. Whenever possible, roentgenographs of the extremities
should be obtained for delineation of skeletal and joint trauma or the
Presence of lesions. Disarticulated limbs should either be frozen or
Preserved in 10% buffered Formalin, and then submitted for age determination
studies. When possible, the cervical spine, consisting of eight mobile
vertebrae should be disarticulated from the head and from the first
thoracic vertebra, which is firmly fixed to the anterior portion of the
carapace. .

VI. Central Nervous System and Organs of Special Sense

The head is then disarticulated. Using a Stryker or hack saw, the
dorsal calvarium of the head can be removed. This requires a transverse
cut through the bone just posterior to the orbits followed by two cuts at
right angles to the first cut through the dorso-lateral calvarium extending
posteriorly to its ending (see Fig. 1). The only structure then holding
the roof of the skull will be the underlying supraoccipital process. This
can be reached with a pair of shears just above the attachments of the
vertebral column to the posterior skull.

When all cuts have been made, the calvarium may be lifted away revealing
three structures: two large lobulated salt glands postorbitally, to each

side, and the cut dorsal surface of the supraoccipital process. If two
incisions are made through the lateral portions of this process, it too
may be lifted away revealing the small, elongate brain. The pineal

gland is just under the roof of the skull dorsal to the pituitary and
located in the middle of a line drawn behind the eyes.

Turning the head upside down, the brain will be held only by cranial
nerves, olfactory tracts and optic nerves which may be carefully cut with
scissors. Examine the brain and fix in 25% Formalin. Section after
fixation. The pituitary, lying below the brain ina bony depression,
is removed with the brain.

The eyes should be removed with curved scissors, cutting within the
orbits and severing the optic nerves, then fixed in 10% Formalin. A



accidently cut. It is, therefore, important that this fluid (note and
record color) be removed frequently by tipping the animal on its side.
If water is available, the cavity may be flushed and drained.

If decomposition has progressed, the fat over the heart and on the
dorso-medial side of the scapula will be black-red and gelatinous.
Further, major organs lose their natural color and appear as if parboiled.
The pleuroperitoneal cavity may contain large amounts of a dark-red
serosanguinous fluid. If these signs are severe, the soft tissues are
unlikely to be suitable for histological examination and need not be
collected. However, information can still be gathered from examination of
gross anatomy and such related parameters as gut contents, parasite load,
toxicology screening and skeletal features.

The basic principles directing the dissection are after those of
Olafson as reported by Van Kruningen (1971):

1. Each part of the carcass is examined in situ. Then it is removed
and examined.

2. Once an organ is removed, it is dissected totally and sections
are taken before continuing the gross dissection.

3. The carcass is examined from outside-in and ventral to dorsal.
1. Throat

A mid-ventral incision is made posteriorly along the neck from the
mandibular symphysis (E, Fig. 1). The skin is reflected, exposing the
underlying trachea and esophagus. The oral cavity is examined, as are
the tongue and glottis. Two incisions (K, L, Fig. 2) are then made along
the inner edges of each mandible through the muscle into the oral cavity.
A finger canbe inserted through the incisions encircling the tongue, and
the tongue pulled upwards after its anterior attachment is severed.

The tongue, larynx, trachea and esophagus are then dissected free of the
neck to a point at which the trachea and esophagus disappear dorsal to

the heart (Fig. 3). The thymic tissue occurs on either side of the trachea
in young animals. Due to the dorsal location of the turtle's lungs, it

is easiest to examine the gastrointestinal system before the respiratory
system.

(1. Gastrointestinal Tract
ass tromtestinal tract

A large, bilobed liver lies over the heart and occupies the anterior
abdominal cavity (Fig. 3). It is dark brown (mahogany), shiny and sharp-
edged under normal conditions. The right lobe is largest and lies ventral
to the small intestine and pancreas. The left lobe covers the anterior
portion of the stomach, which occupies the left abdominal cavity. Grasping
the esophagus and pushing the liver to one side, the stomach can be
dissected free along its inner curvature from the mesentery and liver.

Tie with twine the anterior end of the stomach at its attachment to the
esophagus to prevent the stomach contents from spilling into the body
cavity (N, Fig. 4). Lift the stomach up and cut its esophageal attachment.



abnormal number of barnacles; the muscle masses of the neck and the
extremities are reduced; this may be so marked that the supraoccipital
crest at the back of the head will stand out sharply and the shell appears
too large for the animal. These signs are valid only if the turtle is
relatively fresh and has not undergone advanced decomposition.

The skin and shell are carefully examined for color, texture and
lesions. All lacerations, masses, discolorations, and scars, including
rope burns, tag scars, propeller cuts, and bullet holes, are measured
and described.

The head is carefully inspected for the presence of trauma and
hemorrhage. The eyes are examined for abnormalities with special reference
to the cornea and sclera. The oral cavity and the mucus membranes are
examined for erosions, trauma’and barnacles.

The extremities are then examined for lesions and palpated for fractures.
The anal area is examined for protrusion, exudates, and consistency of
fecal matter. A fecal sample should be placed in a small vial with 10%
Formalin. Necessary scrapings or impression smears of suspected lesions
are made and air dried for submission to the laboratory. Parasites are
removed and placed in AFA solution for later identification (see Clinical
Pathology section).

DISSECTION

Prior to dissection, the turtle is placed on its back. A short
autopsy knife is i.:serted along the bridge between the carapace and the
outermost plastron plates, and the cartilage is cut. The incisions
(A-D, Fig. 1) are made through the skin along the anterior and posterior
plastron edges. The plastron is removed from the underlying muscle masses.
In larger turtles, this is most easily done if the plastron is pulled
upward by an assistant with a hay or stevedore's hook. A mid-ventral
incision (F, Fig. 2) is made between the coracoids and directed posteriorly
to the pelvis. A greyish-green fat overlying the pleuroperitoneum is
normal in Caretta. Incisions (G, H, Fig. 2) are then made along the
lateral border of the muscle masses at the carapace edge and extending
forward so that the muscles and pectoral girdle can be lifted upward and
pushed toward the head, thereby exposing the underlying heart and other
viscera. The muscles may then be removed by disarticulating the contained
girdle at the shoulder (gleno-humoral joint). A similar procedure
(1, J, Fig. 2) can be used to remove the muscle masses of the rear
flippers. A mid-ventral incision (M, Fig. 2) is made separating the
left and right pelvis (in larger turtles long-handled pruning shears
or a saw will be necessary) followed by lifting the pelvis upward and
backward with disarticulation from the femoral heads. In all of these
manipulations, separate the muscle masses carefully from the underlying
pericardium, peritoneum and viscera in a manner that does not disturb the
relationship of organs to one another.

Check the positions of organs (Fig. 3). Frequently the pleuro-
peritoneal cavity will be filled with fluid. This fluid obscures the
lungs and urogenital tracts and may collect in the lungs if they are



ALPHA-NUMERIC CODES USED IN FIGURES

1. Abdominal and Inguinal Fat (green)

2. Pectoral Muscles

3. Pelvic Muscles

4, Tongue

5. Trachea

6. Heart, Ventricle

7. Heart, Auricle

8. Stomach

9. Liver

10. Duodenum

11. Pancreas

12. Intestines

13. Spleen

14. Gall Bladder

15. Lung

16. Mesovarium

17. Kidney

18. Gonad

19. Urinary Bladder

20. Cloaca

A-D. Incision sites for plastron removal.

E. Incision site to expose trachea.

F. Incision site to separate abdominal and pectoral muscle masses.
G-H. Incision sites to allow reflection of pectoral muscle masses.
I-J-M. Incision sites to allow reflection of pelvic muscle masses.
K-L. Incision sites to allow reflection of tongue.

N-0-P. Points of gastrointestional ties to prevent loss of stomach and

intestinal contents.



heart can then be removed from the lungs and examined. Alternatively,
however, if it is easier for the prosector, the heart may be removed from
the lungs while still within the animal (Fig. 4). On the anterior peri-
cardial sac, often obscured by yellow fat, are the thyroid and parathyroid
glands. They are translucent and red but are difficult to see so a wide
excision of the tissue is needed to assure their removal. Just lateral to
these organs and the sac is the Paired pinkish-grey lobular thymus. When
the pericardial sac is cut, be careful to note the amount, consistency
and color of the pericardial fluid. A mid-dorsal opening of the aorta
will expose the aortic valves. The ventricle may be opened by an

incision from its attachment to the right atrium through the apex, to the
left atrium. The arterio-ventricular valves can then be examined and the
incision continued into each atria.

The trachea should be opened and followed to the bronchial bifurcation. -
The lung parenchyma can be examined by following the respiratory tracts
and by transverse incisions through the tissue. Changes in color, texture
and the presence of parasites are noted and sections collected,

A possible, but not certain sign of drowning is a thick, tenacious,
persistent white (or slight pink) foam which can be expressed from
the nostrils by pressure on the throat or is present within the trachea,
bronchi and on cut section of the Tung. In addition, the lungs are greatly
expanded and contain. a watery fluid. When removed and Placed on the
autopsy table, they will collapse and fluid will run from them freely.
Vomitus may be found in the trachea and bronchi. However, with current
limited knowledge, a diagnosis of drowning as the cause of death, based
on gross signs from necropsy, cannot be made.

Iv. Urogenital System and Adrenals

The ovaries lie on each side of the posterior pleuroperitoneal cavity
and can be removed with their unattached oviducts by incising the mesovarium.
Note should be taken of the presence and exact numbers of eggs and degree
of calcification. As the oviducts are followed posteriorly, the urinary
bladder is found just anterior to the cloaca. There are no accessory
cloacal bursae in sea turtles. The bladder,- its cloacal orifice and the
oviductal openings into the cloaca are examined. At this point, the
(ie 2) observed and followed forward to the paired dorsal kidneys

Fig. 5).

By careful dissection, the cloacal opening and rectum can be freed,
pulled upward and forward. Then the attachments of: the urogenital system
and the kidney can be severed and the complete system removed from the
abdominal cavity. The kidneys and bladder should be removed in toto
with the ovary and oviducts, incising just posterior to their cloacal
attachment. The kidneys are dark red to black, covered by a grey-blue
peritoneum and multi-lobed. All organs should be examined for masses,
changes in color or consistency, exudates and Parasites. é

In the male, the cloaca can be opened laterally to expose the penis;
(this organ may be difficult to find in juvenile turtles; sex deter-
mination. in younger animals usually depends‘on histological examination
of gonadal tissue). In either sex, such an incision will expose the orifices



NOAA Technical Memorandum NMFS-SEFC-24

SEA TURTLE NECROPSY MANUAL

Richard E. Wolke and Anita George

December 1981

U.S. DEPARTMENT OF COMMERCE

National Oceanic and Atmospheric Administration
National Marine Fisheries Service

Southeast Fisheries Center

Panama City Laboratory

3500 Delwood Beach Road

Panama City, Florida 32407


Tissue for fixation should be thinly sliced (2 cm x 3 cm x 6 mm) and
not traumatized. Sections should, if possible, contain both normal and
abnormal areas. Sections should be taken from all major organs. This
tissue must never be frozen unless it is being used for virological or
toxicological examination. The best routine fixative is 10% neutral
buffered Formalin (commercial formaldehyde 37-40%, 100 ml; water, 900 ml;
sodium phosphate dibasic anhydrous, 6.5 gms; sodium phosphate monobasic,
4 gms). Two liters is usually sufficient for a single necropsy. Non-
buffered Formalin is acceptable if buffering agents are not available.
Alcohol can be used, but only in an emergency. It tends to make the
tissue quite hard and distorts normal cell shape.

Sections are placed in 10 times their volume of fixative for 24-48
hours. After fixation, sections may be mailed in heat-sealed, polyethylene
food bags with some Formalin-moistened cotton. Make sure that all bottles
and bags are well labeled. Place a waterproof label inside as well as
outside the container. All necropsies should be done as quickly as
possible and certainly within 24 hours of death, if possible (48 hours under
refrigeration). Animals should be removed as quickly as possible from
the water and placed in a cool area.

CLINICAL PATHOLOGY

All fresh tissue and tissue fluids must be submitted to the laboratory
as soon as possible. Blood for hematology and biochemical studies must be
taken before death. Postmortem blood specimens can be used for micro-
biological examination. Tissue for toxicology and virology may be frozen.
Approximately 200 gms of tissue are needed, 15 ml of blood and up to 200 ml
of other fluids. The tissues necessary for toxicological studies are liver,
kidney, brain or spinal cord, fat, skeletal muscle, and eggs if present.
Blood samples should be refrigerated and not frozen. Tissue for microbio-
logical examination may be placed in labeled sterile petri dishes for
submission. Tissues and fluids best suited to determine exposure to
environmental contaminants and toxic substances are liver, muscle, bone,
and stomach contents. Materials to be examined for petroleum derived
hydrocarbons, PCBs, etc. should be stored in glass rather than plastic
containers. .

Before culturing lesions or tissue for bacteria, the bacteriotogical
loop must be flamed and allowed to cool for a few seconds. - The spatula
may be flamed simultaneously and used to sear the surface of the organ
for sterilization. The loop is then passed through the seared area into
the organ proper. The agar is then streaked with the flat surface of
the loop. Be sure to keep the petri dish top over the agar to avoid
contamination. Alternatively a piece of the organ may be removed with
flamed sterile instruments and placed in a sterile petri dish for later
culturing in the laboratory.

Helminths should be relaxed prior to fixation. ‘Trematodes and cestodes
are refrigerated in tap water, and nematodes are relaxed in glacial acetic
acid.

For the routine preservation of parasites AFA solution is excellent
(85% ethyl alcohol, 85 ml; Formalin, 10 ml; acetic acid, 5 ml) or 5%



transverse cut made just posterior to the nostrils and joined by a frontal
cut below the nostrils allows removal of the nares and examination of the
nasal canals. Incise the tympanic membrane over the ear cavity and remove
the slender columella bone which is attached to the skin and extends
medially to the inner ear. It may not be possible to remove the bone in
toto, but even a partial specimen can be valuable for age determination.

A portion of the cervical spinal cord should be removed and preserved.
It is indeed difficult to obtain the whole cord and/or thoracic lumbar
sections which require a ventral approach, hence this tissue is not
obtained routinely.

In general, cause of death of a sea turtle carcass will not be
discernible from gross necropsy observations. Apparent exceptions to this
are obvious signs of trauma, such as bullet wounds, severe lacerations,
massive hemorrhage, and decapitation; but evidence such as this might have
been produced postmortem. The determination of cause of death will require
histopathological examination and analysis in some cases. In other cases,
the determination may be impossible because causes of death have not been
adequately researched.

EQUIPMENT AND FIXATIVES

Equipment will vary from laboratory to laboratory but the following
list is as suggested by Van Kruningen (1971) and. modified (instrument
measurements are in the English system for ease in purchasing):

Rubber gloves

Tissue forceps (6 inch 2 x 3 teeth)

Scalpel (no. 4 with no. 22 blades)

Scissors, straight and curved (6 3/4 inch)

Postmortem shears (7 3/4 inch serrated edges)

Boning knives (5 1/2 inch blade)

Hay or stevedore's hook

Pruning shears

Small hand saw

10. Knife sharpener and steel

11. Stryker electric autopsy saw, hack-saw

12. Bottles, waterproof labels, waterproof. markers

13. Glass microscope slides

14. Sterile petri dishes

15. Blood agar plates; marine agar (Zobell's agar) plates

16. Propane burner

- Spatula and bacteriology loops

18. Blood vials with an anticoagulant (EDTA) (ice and styrofoam
container helpful)

19. Photographic equipment and/or tape recorder

WON DN EWrn—

20. Twine
21. Tape measure and/or calipers
22. Scales

23. 10% and 25% Formalin


At the pylorus, the stomach is cut free and removed after tying both ends
of the transected gut with twine (0, P, Fig. 4). The stomach is then
examined internally after an incision is made along the lesser curvature.
Contents are recorded and preserved in alcohol (40% isopropyl or 70% ethyl).

Before removing the liver, the duodenum is incised mid-ventrally to
expose the bile-duct opening. The duct should be checked for obstructions
by applying pressure to the gall bladder, which lies beneath and within
the back edge of the right liver lobe (Fig. 4). The liver and gall bladder
are removed and carefully examined. The gall bladder is approximately
2 x 3 cm in a 60 cm turtle and dark green. The saccular colon lies
beneath the stomach. (In Dermochelys coriacea, the leatherback turtle;
the arrangement of the anterior gut differs. The stomach lies to the left
and is a U-shaped organ. The esophagus lies along the mid-line dorsally
and extends posteriorly to the anterior edge of the kidneys then turns
and joins the stomach. The pylorus is further forward and the duodenum,
after leaving the stomach, runs backward along the edge of the carapace
on the left.)

The pancreas lies along the duodenum. It is normally grey, irregular
in outline and elongate. Cut the mesentery from the intestine and strip
the intestine from the abdominal cavity.

One or two large mesenteric vessels to the lower intestine should be
removed and fixed for blood fluke. examination in the laboratory. The
intestine is then tied at its entrance to the pelvic inlet, removed from
the cavity and set to one side for further examination. The esophagus,
duodenum, jejunum and large intestine are then opened along the entire
length of the gastrointestinal tract. Lesions are noted and parasites
carefully collected with their numbers and locations recorded.

The mesentery suspénding the intestine is examined.. The pancreas and
the spleen are removed for sectioning. The spleen (Fig. 4) is antedorsal
to the caecum within the mesocolon. It is slate to blue-grey, egg-shaped
and smooth.

Ill. Lungs and Heart

The trachea is a white tube lying ventral to the esophagus and held
open by a series of incomplete cartilaginous rings. It is normally
unobstructed. Just below the heart, it bifurcates to form paired bronchi
which enter the lungs. The paired lungs should be dark pink, partially
air-filled and. of a soft, expansible consistency. On cut section they are
dry. Both lungs are covered by a rather thick greyish-blue sac (pleura).
Their pink color is only apparent after removal of the pleura.

The trachea can be grasped and freed from underlying tissue. While
holding the trachea, the lungs, which lie dorsally against the carapace,
can be dissected free with the heart and great vessels. They should be
removed in toto to simplify identification of the great vessels. The


Tongue and trachea

Muscle masses reflected to reveal viscera.
have been freed and pulled ventrally.

Figure 3.



PREFACE

This manual was prepared by the authors for the National Marine
Fisheries Service under contract no. NA80FACOQ016. Readers are reminded
that taking or possessing sea turtles is prohibited under the Endangered
Species Act of 1973 (PL 93-205) unless proper permits are obtained.
Reference to trade names does not imply endorsement by the National Marine
Fisheries Service.



SPECIMEN COLLECTION CHECKLIST

Skin

External Parasites
Skeletal Muscle
Tongue

Esophagus

Stomach

Liver

Gall Bladder
Pancreas

10. Duodenum

11. Jejunum

12. tleocecal Area

13. Colon

14, Gastrointestinal Parasites
15. Mesenteric Vessels

WON DN EWNH—

16. Spleen
17. Trachea
18. Lung

19. Ventral Pericardial Sac (Thyroid and Parathyroid)
20. Aorta and Great Vessels

21. Atrium

22. Ventricle

23. Gonads and Ducts

24. Urinary Bladder

25. Kidney and Renal Peritoneum (Adrenal)
26. Rectum

27. Cloaca

28.° Salt Glands

29. Brain (whole, sliced lengthwise)

30. Eye

31. Olfactory Block (see text)

32. Scrapings and Smears (where necessary)
33. Front Flipper. and Hind Flipper

34. Cervical Vertebrae

35. Columella

36. Toxicology Screening (frozen about 100 - 200 grams each)
Liver

Kidney

Brain or Spinal Cord

Fat

Skeletal Muscle

Eggs

>rndvanctH

W



Necropsy Report Sheet p. 2-
Head (including mouth, eyes, nares, salt glands, brain)

Gastrointestinal Tract (including liver, spleen, pancreas, mesentery) >

Lungs and Heart (including trachea, bronchi, great vessels)

Urogenital System (including kidneys, bladder, gonads, accessory tubes, cloaca)

Skeletal Muscle and Osseous System



TABLE OF CONTENTS

PREFACE. 2 2. 1 1 we eee ee wee

INTRODUCTION . .. eee we we
EXTERNAL EXAMINATION. rr
DISSECTION . 2... ew ee eee
!. Throat .... wae
ie Gastrointestinal Tract
Ill. Lungs and Heart...

IV. Urogenital System and Adrenals .
V. Connective Tissue, Skeletal Muscle, and Osseous System
Vi. Central Nervous System and Organs of Special Sense

EQUIPMENT AND FIXATIVES .
CLINICAL PATHOLOGY .......
ANAMNESIS AND RECORDS .

DETERMINATION OF CAUSE oF DEATH. .

ACKNOWLEDGMENTS . . . . oe
REFERENCES
SPECIMEN COLLECTION CHECKLIST .
SEA TURTLE NECROPSY REPORT SHEET
PATHOLOGY RECORD .

ALPHA-NUMERIC CODES USED" IN FIGURES

WW ONDA EWWNh HH —



NOAA Technical Memorandum NMFS-SEFC-24

Technical Memorandums are used for documentation
and timely communication of preliminary results,
interim reports, or special-purpose information,
and have not received complete formal review,
editorial control, or detailed editing.

SEA TURTLE NECROPSY MANUAL

Richard E. Wolke and Anita George

Marine Pathology Laboratory

Department of Aquaculture and Pathology
- University of Rhode Island

Kingston, RI 02881

December 1981

U.S. DEPARTMENT OF COMMERCE

Malcolm Baldrige, Secretary

National Oceanic and Atmospheric Administration

John V. Byrne, Administrator

National Marine Fisheries Service

William G. Gordon, Assistant Administrator for Fisheries


——

formol-saline (Formalin, 5 ml; physiologic salt, 95 ml). Relaxation can
be brought about by placing the parasites in warm 70% alcohol and storing
in same.

Scrapings are made with a sharp scalpel and material placed in a
small vial of 10% Formalin. Impression smears are made by cutting across
an organ with a knife or scalpel and pressing (dabbing) the exposed
surface against an alcohol slide. The slide is allowed to air dry and
is submitted in that condition.

ANAMNESIS AND RECORDS

It is imperative that a careful history be taken and that all findings
be recorded. The history should include information about the data and
location of stranding, water temperature, pollutants (oil, mercury), time
and date of death as well as all other pertinent facts.

The prosector must report only what he sees, not what he thinks.
Complete sentences are used and lesions characterized by a number, color,
size, shape, consistency and location.

Photographs are helpful to document descriptions. When the prosector
is alone, it is extremely difficult to record observations and quite easy
to forget those made early in the necropsy. A simple, inexpensive tape
recorder is most helpful in overcoming this problem.

Two pathology report forms are included in this manual. While these
two forms are only samples of the kinds of forms. needed, we recommend
that copies of them be used, unless a better form is available. The Sea
Turtle Necropsy Report Sheet (2 pages) is to be used by the prosector.
The Pathology Record (1 page) is to be used by the pathology laboratory
personnel that report the diagnosis and the cause of death.

DETERMINATION OF CAUSE OF DEATH

A carefully conducted necropsy with complete histopathological
examination may uncover the cause of death in the majority of cases, but

not in all cases. In those instances in which cause of death cannot be
determined, the pathologist can only report morphological changes and
their severity. In all cases, however, the pathologist's final report

should contain a section for morphological diagnosis, etiological
diagnosis, and a statement as to cause of death. Morphological and
etiological diagnoses may be incidental or not of sufficient severity
to have played a role in the turtle's death. Under these circumstances
the pathologist must differentiate between such findings and the actual
cause of death.

The determination of cause of death is dependent, along with other
factors, on the knowledge of diseases and pathological conditions occurring
in that species. Information regarding diseases, their prevalence, and
the clinical manifestations of disease and pathological conditions in sea
turtles is minimal at present. Data are being collected rapidly, but
interpretation often relies on comparisons with similar observations in
higher vertebrates, which may or may not be valid.

9



DATE SEA TURTLE

SUBMITTING ID NO.

NECROPSY REPORT SHEET

; Submitting Prosector Address Phone
Genus Species Carapace LXW Plastron LXW | Head W ] Weight] Sex
Str Str
Curv Curv
HISTORY & CLINICAL SUMMARY: (Water quality; environmental contaminants, stranding location,

date and time of autopsy, etc.)

Specimen submitted | Preservation | Condition of Specimen at Necropsy
Moribund Dead __ Fresh Fixed __
Tissues

Tissues submitted:
Histopathological -
Bacteriological (smears, tissues, cultures) -
Virological (frozen) -

Parasitological (AFA) -
Toxicological (frozen) -

GROSS FINDINGS:
External (including skin, plastron, carapace)